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. 1986 Jun 1;102(6):2310–2317. doi: 10.1083/jcb.102.6.2310

Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes

PMCID: PMC2114259  PMID: 3711147

Abstract

In the developing chick embryo tibia type X collagen is synthesized by chondrocytes from regions of hypertrophy and not by chondrocytes from other regions (Capasso, O., G. Tajana, and R. Cancedda, 1984, Mol. Cell. Biol. 4:1163-1168; Schmid, T. M., and T. F. Linsenmayer, 1985, Dev. Biol. 107:375-381). To investigate further the relationship between differentiation of endochondral chondrocytes and type X collagen synthesis we have developed a novel culture system for chondrocytes from 29-31-stage chick embryo tibiae. At the beginning of the culture these chondrocytes are small and synthesize type II and not type X collagen, but when grown on agarose-coated dishes they further differentiate into hypertrophic chondrocytes that synthesize type X collagen. The synthesis of type X collagen has been monitored in cultured cells by analysis of labeled collagens and in vitro translation of mRNAs. When the freshly dissociated chondrocytes are plated in anchorage-permissive dishes, most of the cells attach and dedifferentiate, as revealed by their fibroblastic morphology. Dedifferentiated chondrocytes, after several passages, can still reexpress the differentiated phenotype and continue their development to hypertrophic, type X collagen-synthesizing chondrocytes. Hypertrophic chondrocytes, when plated in anchorage permissive dishes, attach, maintaining the differentiated phenotype, and continue the synthesis of type X collagen.

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Selected References

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  1. Adams S. L., Alwine J. C., de Crombrugghe B., Pastan I. Use of recombinant plasmids to characterize collagen RNAs in normal and transformed chick embryo fibroblasts. J Biol Chem. 1979 Jun 25;254(12):4935–4938. [PubMed] [Google Scholar]
  2. Adams S. L., Boettiger D., Focht R. J., Holtzer H., Pacifici M. Regulation of the synthesis of extracellular matrix components in chondroblasts transformed by a temperature-sensitive mutant of Rous sarcoma virus. Cell. 1982 Sep;30(2):373–384. doi: 10.1016/0092-8674(82)90235-5. [DOI] [PubMed] [Google Scholar]
  3. Ambesi-Impiombato F. S., Parks L. A., Coon H. G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455–3459. doi: 10.1073/pnas.77.6.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benya P. D., Shaffer J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982 Aug;30(1):215–224. doi: 10.1016/0092-8674(82)90027-7. [DOI] [PubMed] [Google Scholar]
  5. Bonatti S., Cancedda F. D. Posttranslational modifications of Sindbis virus glycoproteins: electrophoretic analysis of pulse-chase-labeled infected cells. J Virol. 1982 Apr;42(1):64–70. doi: 10.1128/jvi.42.1.64-70.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burgeson R. E., Hollister D. W. Collagen heterogeneity in human cartilage: identification of several new collagen chains. Biochem Biophys Res Commun. 1979 Apr 27;87(4):1124–1131. doi: 10.1016/s0006-291x(79)80024-8. [DOI] [PubMed] [Google Scholar]
  7. Cancedda R., Capasso O., Castagnola P., Descalzi-Cancedda F., Quarto N. Deposition of type X collagen in the cartilage extracellular matrix. J Cell Biochem. 1985;28(1):7–14. doi: 10.1002/jcb.240280103. [DOI] [PubMed] [Google Scholar]
  8. Capasso O., Gionti E., Pontarelli G., Ambesi-Impiombato F. S., Nitsch L., Tajana G., Cancedda R. The culture of chick embryo chondrocytes and the control of their differentiated functions in vitro. I. Characterization of the chondrocyte-specific phenotypes. Exp Cell Res. 1982 Nov;142(1):197–206. doi: 10.1016/0014-4827(82)90423-2. [DOI] [PubMed] [Google Scholar]
  9. Capasso O., Quarto N., Descalzi-Cancedda F., Cancedda R. The low molecular weight collagen synthesized by chick tibial chondrocytes is deposited in the extracellular matrix both in culture and in vivo. EMBO J. 1984 Apr;3(4):823–827. doi: 10.1002/j.1460-2075.1984.tb01891.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Capasso O., Tajana G., Cancedda R. Location of 64K collagen producer chondrocytes in developing chicken embryo tibiae. Mol Cell Biol. 1984 Jun;4(6):1163–1168. doi: 10.1128/mcb.4.6.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Caplan A. I. Effects of the nicotinamide-sensitive teratogen3-acetylpyridine on chick limb cells in culture. Exp Cell Res. 1970 Oct;62(2):341–355. doi: 10.1016/0014-4827(70)90564-1. [DOI] [PubMed] [Google Scholar]
  12. Chang H. C., Jones O. W., Masui H. Human amniotic fluid cells grown in a hormone-supplemented medium: suitability for prenatal diagnosis. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4795–4799. doi: 10.1073/pnas.79.15.4795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Coon H. G. Clonal stability and phenotypic expression of chick cartilage cells in vitro. Proc Natl Acad Sci U S A. 1966 Jan;55(1):66–73. doi: 10.1073/pnas.55.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gibson G. J., Beaumont B. W., Flint M. H. Synthesis of a low molecular weight collagen by chondrocytes from the presumptive calcification region of the embryonic chick sterna: the influence of culture with collagen gels. J Cell Biol. 1984 Jul;99(1 Pt 1):208–216. doi: 10.1083/jcb.99.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gibson G. J., Flint M. H. Type X collagen synthesis by chick sternal cartilage and its relationship to endochondral development. J Cell Biol. 1985 Jul;101(1):277–284. doi: 10.1083/jcb.101.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibson G. J., Kielty C. M., Garner C., Schor S. L., Grant M. E. Identification and partial characterization of three low-molecular-weight collagenous polypeptides synthesized by chondrocytes cultured within collagen gels in the absence and in the presence of fibronectin. Biochem J. 1983 May 1;211(2):417–426. doi: 10.1042/bj2110417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gibson G. J., Schor S. L., Grant M. E. Effects of matrix macromolecules on chondrocyte gene expression: synthesis of a low molecular weight collagen species by cells cultured within collagen gels. J Cell Biol. 1982 Jun;93(3):767–774. doi: 10.1083/jcb.93.3.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gionti E., Capasso O., Cancedda R. The culture of chick embryo chondrocytes and the control of their differentiated functions in vitro. Transformation by rous sarcoma virus induces a switch in the collagen type synthesis and enhances fibronectin expression. J Biol Chem. 1983 Jun 10;258(11):7190–7194. [PubMed] [Google Scholar]
  19. Kielty C. M., Kwan A. P., Holmes D. F., Schor S. L., Grant M. E. Type X collagen, a product of hypertrophic chondrocytes. Biochem J. 1985 Apr 15;227(2):545–554. doi: 10.1042/bj2270545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Levitt D., Dorfman A. The irreversible inhibition of differentiation of limb-bud mesenchyme by bromodeoxyuridine. Proc Natl Acad Sci U S A. 1972 May;69(5):1253–1257. doi: 10.1073/pnas.69.5.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lowe M. E., Pacifici M., Holtzer H. Effects of phorbol-12-myristate-13-acetate on the phenotypic program of cultured chondroblasts and fibroblasts. Cancer Res. 1978 Aug;38(8):2350–2356. [PubMed] [Google Scholar]
  23. Mayne R., Elrod B. W., Mayne P. M., Sanderson R. D., Linsenmayer T. F. Changes in the synthesis of minor cartilage collagens after growth of chick chondrocytes in 5-bromo-2'-deoxyuridine or to senescence. Exp Cell Res. 1984 Mar;151(1):171–182. doi: 10.1016/0014-4827(84)90366-5. [DOI] [PubMed] [Google Scholar]
  24. Mayne R., Vail M. S., Mayne P. M., Miller E. J. Changes in type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity. Proc Natl Acad Sci U S A. 1976 May;73(5):1674–1678. doi: 10.1073/pnas.73.5.1674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mayne R., Vail M. S., Miller E. J. The effect of embryo extract on the types of collagen synthesized by cultured chick chondrocytes. Dev Biol. 1976 Dec;54(2):230–240. doi: 10.1016/0012-1606(76)90301-8. [DOI] [PubMed] [Google Scholar]
  26. Miller E. J. Isolation and characterization of a collagen from chick cartilage containing three identical alpha chains. Biochemistry. 1971 Apr 27;10(9):1652–1659. doi: 10.1021/bi00785a024. [DOI] [PubMed] [Google Scholar]
  27. Pacifici M., Holtzer H. Effects of a tumor-promoting agent on chondrogenesis. Am J Anat. 1977 Sep;150(1):207–212. doi: 10.1002/aja.1001500116. [DOI] [PubMed] [Google Scholar]
  28. Quarto N., Cancedda R., Descalzi-Cancedda F. Purification and characterization of the low-molecular-mass (type X) collagen from chick-embryo tibial cartilage. Eur J Biochem. 1985 Mar 1;147(2):397–400. doi: 10.1111/j.1432-1033.1985.tb08763.x. [DOI] [PubMed] [Google Scholar]
  29. Reese C. A., Mayne R. Minor collagens of chicken hyaline cartilage. Biochemistry. 1981 Sep 15;20(19):5443–5448. doi: 10.1021/bi00522a014. [DOI] [PubMed] [Google Scholar]
  30. Reese C. A., Wiedemann H., Kühn K., Mayne R. Characterization of a highly soluble collagenous molecule isolated from chicken hyaline cartilage. Biochemistry. 1982 Mar 2;21(5):826–830. doi: 10.1021/bi00534a002. [DOI] [PubMed] [Google Scholar]
  31. Remington M. C., Bashey R. I., Brighton C. T., Jimenez S. A. Biosynthesis of a disulphide-bonded short-chain collagen by calf growth-plate cartilage. Biochem J. 1984 Nov 15;224(1):227–233. doi: 10.1042/bj2240227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schmid T. M., Conrad H. E. A unique low molecular weight collagen secreted by cultured chick embryo chondrocytes. J Biol Chem. 1982 Oct 25;257(20):12444–12450. [PubMed] [Google Scholar]
  33. Schmid T. M., Linsenmayer T. F. A short chain (pro)collagen from aged endochondral chondrocytes. Biochemical characterization. J Biol Chem. 1983 Aug 10;258(15):9504–9509. [PubMed] [Google Scholar]
  34. Schmid T. M., Linsenmayer T. F. Developmental acquisition of type X collagen in the embryonic chick tibiotarsus. Dev Biol. 1985 Feb;107(2):373–381. doi: 10.1016/0012-1606(85)90319-7. [DOI] [PubMed] [Google Scholar]
  35. Schmid T. M., Linsenmayer T. F. Immunohistochemical localization of short chain cartilage collagen (type X) in avian tissues. J Cell Biol. 1985 Feb;100(2):598–605. doi: 10.1083/jcb.100.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scott-Savage P., Hall B. K. The timing of the onset of osteogenesis in the tibia of the embryonic chick. J Morphol. 1979 Dec;162(3):453–463. doi: 10.1002/jmor.1051620310. [DOI] [PubMed] [Google Scholar]
  37. Solursh M., Linsenmayer T. F., Jensen K. L. Chondrogenesis from single limb mesenchyme cells. Dev Biol. 1982 Nov;94(1):259–264. doi: 10.1016/0012-1606(82)90090-2. [DOI] [PubMed] [Google Scholar]
  38. Solursh M., Reiter R. S. Determination of limb bud chondrocytes during a transient block of the cell cycle. Cell Differ. 1975 Jun;4(3):131–137. doi: 10.1016/0045-6039(75)90034-2. [DOI] [PubMed] [Google Scholar]
  39. Stocum D. L., Davis R. M., Leger M., Conrad H. E. Development of the tibiotarsus in the chick embryo: biosynthetic activities of histologically distinct regions. J Embryol Exp Morphol. 1979 Dec;54:155–170. [PubMed] [Google Scholar]
  40. West C. M., Lanza R., Rosenbloom J., Lowe M., Holtzer H., Avdalovic N. Fibronectin alters the phenotypic properties of cultured chick embryo chondroblasts. Cell. 1979 Jul;17(3):491–501. doi: 10.1016/0092-8674(79)90257-5. [DOI] [PubMed] [Google Scholar]
  41. Yoshimura M., Jimenez S. A., Kaji A. Effects of viral transformation on synthesis and secretion of collagen and fibronectin-like molecules by embryonic chick chondrocytes in culture. J Biol Chem. 1981 Sep 10;256(17):9111–9117. [PubMed] [Google Scholar]
  42. Zanetti N. C., Solursh M. Induction of chondrogenesis in limb mesenchymal cultures by disruption of the actin cytoskeleton. J Cell Biol. 1984 Jul;99(1 Pt 1):115–123. doi: 10.1083/jcb.99.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. von der Mark K., van Menxel M., Wiedemann H. Isolation and characterization of new collagens from chick cartilage. Eur J Biochem. 1982 May;124(1):57–62. doi: 10.1111/j.1432-1033.1982.tb05905.x. [DOI] [PubMed] [Google Scholar]

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